We construct the Seiberg-Witten curve for the E-string theory in six-dimensions. The curve is expressed in terms of affine E_8 characters up to level 6 and is determined by using the mirror-type transformation so that it reproduces the number of holomorphic curves in the Calabi-Yau manifold and the amplitudes of N=4 U(n) Yang-Mills theory on 1/2 K3. We also show that our curve flows to known five- and four-dimensional Seiberg-Witten curves in suitable limits.